Study Of Mechanism And AMACR Overexpression Promote Hepatocellular Carcinoma Cells Proliferation

Objective: AMACR is a cytoplasmic protein, mainly present in human peroxisomes and mitochondria, β oxidation of fatty acids and fatty acid derivatives involved in a side chain and play an important role in the oxidation of bile acid metabolic intermediates, and its high expression of the proliferation of tumor has a positive role in promoting, but this finding has not been confirmed in liver cancer. The group will use the label-free quantitative proteomics technology to analysis the Influence of AMACR for Hepatocellular Carcinoma cell growth and proliferation.Methods: we recombined the AMACR-overexpressed cell line by transducting the Lenti-AMACR plasmid into HepG2 cell. To detect the influence of AMACR for Hepatocellular Carcinoma cell proliferation by CCK-8 assay.Followed by the extraction of the whole protein from AMACR-overexpressed HepG2 cells. Afterwards, the samples was identified and quantified by label-free quantitative proteomic approche. The identified proteins were selected differentially expressed proteins by statistical methods. subsequently analyzed by bioinformatics, and verified by qRT-PCR.Results: On the basis of successful construction of AMACR-overexpressed cell line, CCK-8 assay reveal AMACR can promote Hepatocellular Carcinoma cell growth and proliferation. the whole protein isolated from AMACR-overexpressed cell line was comparatively quantified by 2D-LC MS/MS. A total of 138 differentially expressed proteins(including 124 up-regulated proteins and 14 down-regulated proteins)were filtered out. These dysregulated proteins were majorly involved into metabolic process and cellular process, which suggested a big change of biological behaviors occurred in the host cell due to AMACR overexpression. Further analysis of dysregulated proteins involved signalings revealed that AMACR modify the host cell mainly through modulating the ERK1/2 and NF-κB signalings. The expression of AMACR and FABP5 at mRNA level consist with MS results.Conclusions: AMACR can enhance the proliferation of HCC through modulating the ERK1/2 and NF-κB signalings and up-regualting the expression of some carcinogenic factors.